Water Soluble Vitamins

D. A. Ballesteros, MD, MBAH De La Salle Health Sciences Institute College of Medicine Department of Biochemistry

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Merciful and loving Father, our true source of light and wisdom, our only source of health and healing as you called us to care for our brethren help us to see You in each of our classmates, future patients, their families and our teachers. Help us to understand that we serve as instruments by which you heal and bring comfort. Great Healer, please give us a heart that listens carefully and patiently to others, Help us to recognize the needs of each and give us wisdom, willingness, strength, courage and resources to meet these needs, according to Your will. Give us a smile and a gentle sense of humor and help us lighten the trying times. May we learn something new each day, not getting lost in our daily routine, and help us to persist in our commitment, vocation and calling. Amen.

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Introduction
Vitamine vita (life) and amine
Casimir Funk, 1911

Vitamins
Organic nutrients Required in the diet Needed only in minute amount

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Classification
Fat soluble vitamins
Function as coenzymes, hormones and antioxidants

Water soluble vitamins

Precursor for coenzymes and antioxidants

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Classification
Water soluble vitamins
Dissolve in water Not stored extensively (except Vitamin B12) Eliminated in the urine Easily destroyed or washed out during storage and preparation

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Characteristics
Diverse physical properties Active only in COENZYME form

NOT SOURCE OF ENERGY NOT FOR STRUCTURAL PURPOSE

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Role as Coenzymes
CARRIERS of chemical moieties in chemical reactions

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Water Soluble Vitamins

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Thiamine

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Structure
Thiamine THIAZOLE

PYRIMIDINE

Methylene bridge

TPP

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Absorption, Transport and Metabolism

Absorbed in the SI by a carrier mediated transport (intake less than 5mg/day) Higher intake - passive diffusion Phosphorylation in the jejunal mucosa TPP Half of the body stores: skeletal muscles Thiamine and its catabolites are excreted in the urine
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Functions
Required mainly for CHO metabolism Coenzyme for the following enzymes:
Pyruvate dehydrogenase (pyrAcetylCoA) -ketoglutarate dehydrogenase (-KGSuccinylCoA) Keto-acid dehydrogenase and leu, ile, val metab Transketolase (Pentose Phosphate Pathway)
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Nutritional Requirements
0.4mg/1000kcal related to energy metabolism RDA of 1.0 to 1.5 mg for adults
Increased with increased muscular activity, dietary carbohydrates and in pregnancy and lactation

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Deficiency Manifestations
Appears in:
Caloric or carbohydrate intake disproportionately high compared to thiamine intake Chronic alcoholics

Overall picture icludes neurologic, cardiovascular and gastrointestinal disorders referred to as beriberi
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Deficiency Manifestations
Dry beriberi (peripheral neuritis)
Poor appetite, peripheral neuropathy, fatigue, muscle pain and weakness

Wet beriberi (cardiac manifestation)

Neurologic and cardiovascular

Cerebral beriberi (Wernicke-Korsakoffs syndrome) Infantile beriberi

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Deficiency Manifestations
Cerebral beriberi (Wernicke-Korsakoffs syndrome)
Mental confusion, ataxia, diplopia, nystagmus, anorexia, edema, weakness and paralysis

Infantile beriberi

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Thiamine Assay
Whole blood or Erythrocyte transketolase Reference Interval
Whole blood = 9-12 umol/hr/mL (150-200U/L) Erythrocyte = 0.75 to 1.30 U/g of hgb

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Antimetabolites
Thiamine can be destroyed if the diet contains thiaminase Thiaminase is present in raw fish and seafood

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Riboflavin

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Structure
Ribitol

Isoalloxazine ring

Heat stable, photosensitive

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Absorption, Transport and Metabolism

Ingested in the form of flavoproteins FAD and FMN components are released from protein complex in the stomach, free riboflavin is released in the intestine Conversion to coenzyme occurs within the cellular cytoplasm (small intestine, liver, heart and kidney) Main storage form is FAD
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Active/Coenzyme Forms
Flavin Mononucleotide (FMN) Flavin adenine dinucleotide (FAD)

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VIT. B2 CONVERSION TO COFACTORS

Intestinal mucosal cells

RIBOFLAVIN
ATP ADP

FMN
ATP

FAD

PPi

liver

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Functions
Precursor of FMN and FAD, which are required by several redox reactions in metabolism Maintenance of mucosal epithelial and ocular tissues Involved in metabolism of iron, pyridoxine and folate Involved in protection against peroxidation in metabolism of xenobiotics
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FMN-FAD requiring enzymes

Flavoprotein enzyme -amino acid oxidase Xanthine oxidase Mitochondrial glycerol-3-PO4 dehydrogenase Succinate dehydrogenase Acyl-CoA dehydrogenase NADH dehydrogenase Pyruvate dehydrogenase and -KG dehydrogenase Pathway/reaction Deamination of amino acids Purine degradation Transfer of reducing equivalents from cytosol into mitochondria Citric acid cycle Fatty acid oxidation Respiratory chain Oxidative decarboxylation of pyr and -KG

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Nutritional Requirements
RDA 1.3 to 1.7 mg for adults Related to protein use and increases during growth, pregnancy, lactation and wound healing

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Deficiency Manifestations
Can be seen in the following conditions:
Malabsorption, malnutrition, anorexia, chronic alcoholism and drug intake (barbiturate)

Avitaminosis is characterized by:

Angular stomatitis, cheilosis, glossitis, seborrheic dermatitis, vascularization of the cornea

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Riboflavin Assay
FAD dependent glutathione reductase Reference interval
Flurometric method (erythrocyte) = 10 to 15 ug/dL (266 to 1330 nmol/L) Serum /plasma = 4 to 24 ug/dL (106 to 638 nmol/L)

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Niacin

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Structure

Niacin is a general name for nicotinic acid and nicotinamide Simple derivatives of pyridine

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Structure
Nicotinamide Adenine Dinucleotide (NAD+)

Nicotinamide

Adenine

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Absorption, Transport and Metabolism

Both acid and amide forms are absorbed readily Converted to coenzyme forms in the blood, kidney, brain and liver Humans excrete 1-methylnicotinamide and 1-methyl-3-carboxamido-6-pyridone as primary urinary metabolites

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Active forms
Nicotinamide adenine dinucleotide (NAD+) Nicotinamide adenine dinucleotide phosphate (NADP+)

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Functions
NAD+ and NADP+ are involved in a variety of redox reactions catalyzed by dehydrogenases
NAD+ oxidative pathways NADP + reductive synthesis

May have a role in DNA repair and other cellular responses to DNA damage

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Nutritional Requirement
RDA 15 to 20 mg Trp can provide about 10% of the total niacin requirement

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Deficiency involves the skin, GI tract and CNS Dermatitis Diarrhea Dementia
Vitamin B6 pyridoxal phosphate coenzyme in niacin to tryptophan

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Structure

-alanine

Pantoic acid

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Absorption, Transport and Metabolism

CoA, the form in which pantothenic acid is ingested is hydrolyzed by intestinal enzymes to pantetheine and absorbed in into the portal circulation About 80% in the tissues is in CoA form B-mercaptoethylamine excreted in the urine

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Active forms
Coenzyme-A (CoA) Acyl carrier protein (ACP)

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Function
The thiol (-SH) group of CoA and ACP acts as carrier of acyl groups CoA participates in TCA, FA metabolism, acetylation of drugs, synthesis of cholesterol and utilization of ketone bodies ACP participate in FA synthesis reactions

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Nutrional Requirement
No RDA established No clear-cut case of pantothenic acid deficiency reported
Paresthesias, headache, dizziness and GI malfunction Burning feet syndrome (malnourished POW in the 1940s)

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Pyridoxine

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Structure

Pyridoxamine Pyridoxal

Equal vitamin activity Can be interconverted

Pyridoxine

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Active Form
Pyridoxal phosphate
formed from phosphorylation of all three forms of Vit B6

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Absorption, Transport and Metabolism

Vitamers are released by dephosphorylation from their phosphate esters by intestinal alkaline phosphatase Readily absorbed by intestinal mucosal cells, with cytoplasmic pyridoxal kinase rephosphorylating them FMN dependent oxidase catalyze conversion to Pyridoxal Phosphate
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Functions
Concentration is high in the brain, liver and muscle Associated with muscle glycogen phsophorylase Coenzyme for AA metabolism (aminotransferase, amino acid decarboxylase Deamination of ser, thr and cys
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Function
Conversion of trp to serotonin and trp to niacin Production of heme aminolevulinic acid synthetase Important in steroid hormone action
Removes hormone-receptor complex from DNA binding

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Deficiency Manifestations
Microcytic hypochromic anemia Peripheral neuropathy

Drug antagonism (INH and penicillamide) Alcoholism (hydrolysis of PO4 from PP)

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Vitamin B6 Assay
Blood transaminases Reference Interval (plasma) 5-30ng/mL or 20 to 121 nmol/L

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BIOTIN
Formerly known as

Vitamin H

Liver, kidney, vegetable and egg yolk Synthesized by intestinal bacteria

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Structure
Imidazole derivative consisting of an tetrahydrothiophene ring bound to imidazole ring and valeric acid

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Absorption, Transport and Metabolism

Absorbed readily Cleared from the circulation and taken up by the muscles and liver

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Reference Interval
Whole blood or serum: 0.82 to 2.05 nmol/L Deficiency manifestation unknown
May occur in long term TPN Intake of large amounts of uncooked eggs avidin (binds the imidazole group of biotin and prevents biotin absorption) Use of antibiotics
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Functions
Coenzyme of carboxylase reactions (carrier of CO2)
Acetyl CoA carboxylase (FA synthesis) Pyruvate carboxylase (gluconeogenesis) Propionyl CoA carboxylase (propinate to succinate) -methyl crotonyl CoA carboxylase (branched chain AA catabolism)

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Folic Acid

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Structure
Family of compounds related to pteroic acid

Pteridine derivative

PABA

Glutamic acid

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Active Form
Tetrahydrofolate (THF)

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Absorption, Transport and Metabolism

Polyglutamate forms are converted to monoglutamates by glutamyl hydrolase in the intestinal mucosa After cellular uptake, it is reduced and methylated

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Function
Acts as carrier of one carbon moieties
(through N5 and N10 position/s)

Major reactions involve conversion

of ser to gly Synthesis of thmidilate Catabolism of histidine Synthesis of purine Synthesis of met from homocysteine
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Nutritional Requirement
RDA 200 mg

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Deficiency Manifestation
Magaloblastic anemia Sensory loss Neuropsychiatric changes Accumulation of FIGLU in the urine
Formiminoglutamate from histidine

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Cobalamin

Absent in plant products Human obtain small amounts from intestinal flora

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Structure

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Active Form
Methylcobalamin Deoxyadenosylcobalamin

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Absorption and Storage

Bound to hapcorrin, a salivary protein, in the upper GI Binds to intrinsic factor (from parietal cells) The only water soluble vitamin that is stored in significant amount in the liver and bone marrow Transport in plasma
Hapcorrin (transcobalamin I) and transcobalamin II
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Function
Vitamin B12 dependent enzymes
Methylmalonyl CoA mutase Leucine aminomutase Methionine synthase

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Nutritional Requirement
RDA 400ug

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Deficiency Manifestations
Megaloblastic anemia Neuropathy

Common causes
Pernicious anemia (deficiency of intrinsic factor) Atrophic gastritis Strict vegans
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Reference Interval
Fasting state 200 to 835pg/mL (148 to 620pmol/L)

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Ascorbic Acid

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Structure
Six carbon sugar derivative

Human cannot synthesize ascorbic acid due to lack of gluconolactone oxidase

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Functions
Function as a reducing agent
Collagen biosynthesis
Hydroxylation of pro and lys

Steroid synthesis
Hydroxylation of steroids

Carnithine synthesis
Hydroxylation of -butyrobetain to carnitine

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Functions
Microsomal drug metabolism Bile acid formation
7-hydroxylase step

Degradation of tyrosine
Oxidation ofp-hydrophenylpyruvate to homogentisate

Folate metabolism
Conversion of folic acid to THF

Absorption of iron
Reduce iron to the ferrous state

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Functions
Water soluble antioxidant
Reduces oxidized tocopherol produced in the free radical scavenging by Vit E Prevents oxidation of LDL Prevention of cancer by inhibiting nitrosamine formation

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Absorption, Transport and Metabolism

Absorbed in the stomach, some converted to dehydro form Physiologic pH, unchanged dehydroascorbic acid passes cell membrane faster than L-ascorbate Highest concentration in the adrenals, pituitary and retina

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Reference Interval
Plasma 0.4 to 1.5 mg/dL Nutritional requirements
60 mg adults 70mg pregnant women 95 mg lactating women 100 mg smokers

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